Chemical and biological differences between original and mimetic pentosan polysulfates.

Pentosan polysulfate sodium (PPS) is a semi-synthetic, heparin-like polysaccharide with manifold therapeutic actions. It is approved for treatment of bladder pain syndrome / interstitial cystitis in humans and treatment of musculoskeletal diseases in animals. PPS is produced by a complex procedure using beech wood as starting material. It consists of a mixture of sulfated glucuronoxylans, whose structural composition cannot be fully characterized by physicochemical analysis. The question arises whether PPS follow-on products are identical with the original and thus meet the requirement for generic drug application. The aim of this study was to investigate whether commercially available PPS products differ in physicochemical characteristics and biological effects from the original. Ten PPS preparations from different manufactures were analyzed using orthogonal analytical techniques including, inter alia, size exclusion chromatography with triple detection, nuclear magnetic resonance spectroscopy, and high-resolution mid-infrared spectroscopy in aqueous solution with chemometric evaluation. For functional analysis, we measured the plasma kallikrein generation in human plasma and FXII activation. The study revealed significant structural and biological differences between PPS from different sources. Therefore, follow-on products cannot be considered identical but at best similar to original PPS. However, their similar efficacy and safety have still to be proven by comprehensive studies.

Pentosan Polysulfate Inhibits Attachment and Infection by SARS-CoV-2 In Vitro: Insights into Structural Requirements for Binding.

Two years since the outbreak of the novel coronavirus SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) pandemic, there remain few clinically effective drugs to complement vaccines. One is the anticoagulant, heparin, which in 2004 was found able to inhibit invasion of SARS-CoV (CoV-1) and which has been employed during the current pandemic to prevent thromboembolic complications and moderate potentially damaging inflammation. Heparin has also been shown experimentally to inhibit SARS-CoV-2 attachment and infection in susceptible cells. At high therapeutic doses however, heparin increases the risk of bleeding and prolonged use can cause heparin-induced thrombocytopenia, a serious side effect. One alternative, with structural similarities to heparin, is the plant-derived, semi-synthetic polysaccharide, pentosan polysulfate (PPS). PPS is an established drug for the oral treatment of interstitial cystitis, is well-tolerated, and exhibits weaker anticoagulant effects than heparin. In an established Vero cell model, PPS and its fractions of varying molecular weights inhibited invasion by SARS-CoV-2. Intact PPS and its size-defined fractions were characterized by molecular weight distribution and chemical structure using nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry, then employed to explore the structural basis of interactions with SARS-CoV-2 spike protein receptor-binding domain (S1 RBD) and the inhibition of Vero cell invasion. PPS was as effective as unfractionated heparin, but more effective in inhibiting cell infection than low-molecular-weight heparin (on a weight/volume basis). Isothermal titration calorimetry and viral plaque-forming assays demonstrated size-dependent binding to S1 RBD and inhibition of Vero cell invasion, suggesting the potential application of PPS as a novel inhibitor of SARS-CoV-2 infection.

Combined quality of life and posttraumatic growth evaluation during follow-up care of patients suffering from oral squamous cell carcinoma.

Oral cancer therapy is associated with a loss in health-related quality of life (HRQOL) and can also lead to post-traumatic growth (PTG). The current study analyzed the relationship between HRQOL, PTG and influencing clinical factors after treatment. The coherent clinical data of 15 patients were retrospectively analyzed over a 1-year study period. HRQOL and PTG were studied using the University of Washington Quality of Life Version 4 (UW-QOL v4) and Posttraumatic Growth Inventory (PTGI) questionnaires. The results revealed that HRQOL was significantly decreased in a pre- to postoperative manner (P=0.011). Sex demonstrated a nearly significant effect on HRQOL (P=0.058). PTG was experienced the most after surgery, and continuously decreased over the 1-year study period. Patient age had a significant effect on PTG (P=0.040). A significant correlation was also established between HRQOL and PTG (P<0.05). HRQOL and PTG are important influencing factors during postoperative tumor follow-up care and should be simultaneously recorded to address individual patient needs and improve quality of treatment.

Serum uromodulin-a marker of kidney function and renal parenchymal integrity.

Background: An ELISA to analyse uromodulin in human serum (sUmod) was developed, validated and tested for clinical applications. Methods: We assessed sUmod, a very stable antigen, in controls, patients with chronic kidney disease (CKD) stages 1-5, persons with autoimmune kidney diseases and recipients of a renal allograft by ELISA. Results: Median sUmod in 190 blood donors was 207 ng/mL (women: men, median 230 versus 188 ng/mL, P = 0.006). sUmod levels in 443 children were 193 ng/mL (median). sUmod was correlated with cystatin C (rs = -0.862), creatinine (rs = -0.802), blood urea nitrogen (BUN) (rs = -0.645) and estimated glomerular filtration rate (eGFR)-cystatin C (rs = 0.862). sUmod was lower in systemic lupus erythematosus-nephritis (median 101 ng/mL), phospholipase-A2 receptor- positive glomerulonephritis (median 83 ng/mL) and anti-glomerular basement membrane positive pulmorenal syndromes (median 37 ng/mL). Declining sUmod concentrations paralleled the loss of kidney function in 165 patients with CKD stages 1-5 with prominent changes in sUmod within the 'creatinine blind range' (71-106 µmol/L). Receiver-operating characteristic analysis between non-CKD and CKD-1 was superior for sUmod (AUC 0.90) compared with eGFR (AUC 0.39), cystatin C (AUC 0.39) and creatinine (AUC 0.27). sUmod rapidly recovered from 0 to 62 ng/mL (median) after renal transplantation in cases with immediate graft function and remained low in delayed graft function (21 ng/mL, median; day 5-9: relative risk 1.5-2.9, odds ratio 1.5-6.4). Immunogold labelling disclosed that Umod is transferred within cytoplasmic vesicles to both the apical and basolateral plasma membrane. Umod revealed a disturbed intracellular location in kidney injury. Conclusions: We conclude that sUmod is a novel sensitive kidney-specific biomarker linked to the structural integrity of the distal nephron and to renal function.

Osteoblast-like cells with different embryologic origins behave differently in increasing zoledronic acid concentrations: a pilot study in pigs.

OBJECTIVE: We compared the intraindividual effects of increasing zoledronic acid (ZA) concentrations on osteoblast-like cells with different embryologic origins. STUDY DESIGN: Cultured osteoblast-like cells from mandible and iliac crest bone samples of domestic pigs were exposed to increasing concentrations of ZA (0, 10-8, 10-6, and 10-4 M). Proliferation was assessed by cell counting. Receptor activator of nuclear factor kB ligand and osteoprotegerin (OPG) messenger RNA expression were assessed at 0, 1, 4, 7, and 10 days. RESULTS: The OPG expression level was higher in the iliac crest than in the mandible. Neither ZA concentration nor the cells' origin affected the expression of receptor activator of nuclear factor kB ligand. At 10-6 M, OPG expression from both locations reached the same level after 7 days of cultivation, as OPG expression increased to a greater extent in the mandible in comparison to the iliac crest. CONCLUSION: Cultured mandibular osteoblast-like cells reacted more sensitively to high ZA concentrations than did osteoblast-like cells from the iliac crest.

Primary stability and osseointegration of dental implants in polylactide modified bone - A pilot study in Goettingen minipigs.

The present study aimed to evaluate primary stability (PS) and osseointegration of dental implants in polylactide [70/30 poly(l-lactide-co-d, l-lactide); (PLDLA)] modified bone in 30 Goettingen minipigs. Each animal received three implants per jaw quadrant. In a split-mouth design, one side of the maxilla and mandible was randomly allocated to the experimental treatment (PLDLA applied into the drill hole before implantation), while the contralateral sides served as intraindividual controls (no PLDLA applied). The required insertion torque and the implant stability quotient (ISQ) were measured during implantation. ISQ, volume density (VD) of new bone formation (NBF), and the bone-implant contact (BIC) were evaluated at the end of the observation period (1, 3, 6, 12, and 24 months, respectively) in six animals each. Across all study groups, the PLDLA treatment resulted in a) a comparable insertion torque, b) an equivalent ISQ, c) a reduced BIC, and d) a reduced VD of NBF, as opposed to the untreated controls. In conclusion, the PLDLA treatment did not affect the PS, but rather led to an impaired osseointegration, which was particularly strong in the compact mandibular bone, and decreased in the spongious maxillary bone. PLDLA induced anchoring in spongious bone should be evaluated in further investigations.

Connexin subtype expression during oral carcinogenesis: A pilot study in patients with oral squamous cell carcinoma.

Gap junctional intercellular communication (GJIC) and connexin (Cx) expression were reported in association with carcinogenesis in various types of tumours. In an earlier histomorphometric study, the protein levels of Cx subtypes 26, 43 and 45 were differentially expressed in oral squamous cell carcinoma (OSCC), corresponding lymph node metastases and dysplasia-free oral mucosa. Moreover, membrane Cx43 acted as an independent prognostic marker in OSCC tissues. This study aimed to confirm the expression of described Cx subtypes at the mRNA level. Hence, a reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis of Cx26, Cx43 and Cx45 gene expressions was performed in paired carcinoma and mucosa samples of 15 OSCC patients. Additionally, we assessed the interaction between Cx subtype expression and clinicopathological routine parameters. The RT-qPCR analysis revealed that Cx26 was downregulated in OSCC (P=0.01), while Cx43 was marginally upregulated in cancer tissue (P=0.04). Cx45 was significantly overexpressed in OSCC tissue compared with the intraoral mucosa controls (P<0.01), and remained unchanged at different tumour stages. No significant interactions between differential Cx subtype expression and clinicopathological routine parameters were observed. In conclusion, Cx regulation at the transcriptional level appears to be an early event during the initiation and development of OSCC, and is maintained during further progression. However, the mRNA-protein correlation is variable. This may be indicative of post-transcriptional, translational and degradation regulations being associated with the determination of Cx protein concentration during oral carcinogenesis.

Reduced somatosensory impairment by piezosurgery during orthognathic surgery of the mandible.

PURPOSE: This clinical trial aimed to test the hypothesis that piezosurgery causes reduced nerval irritations and, thus, reduced somatosensory impairment when used in orthognathic surgery of the mandible. METHODS: To this end, 37 consecutive patients with Angle Class II and III malocclusion were treated using bilateral sagittal split osteotomies (BSSO) of the mandible. In a split mouth design, randomized one side of the mandible was operated using a conventional saw, while a piezosurgery device was used on the contralateral side. In order to test the individual qualities of somatosensory function, quantitative sensory testings (QSTs) were performed 1 month, 6 months and 1 year after surgery. RESULTS: A comparison of the data using a two-way analysis of variance (ANOVA) revealed a significant reduction in postoperative impairment in warm detection threshold (WDT) (P = 0.046), a decreased dynamic mechanical allodynia (ALL) (P = 0.002) and a decreased vibration detection threshold (VDT) (P = 0.030) on the piezosurgery side of the mandible as opposed to the conventionally operated control side. In the remaining QSTs, minor deviations from the preoperative baseline conditions and a more rapid regression could be observed. CONCLUSIONS: Piezosurgery caused reduced somatosensory impairment and a faster recovery of somatosensory functions in the present investigation.

Mandibular reconstruction using a calcium phosphate/polyethylene glycol hydrogel carrier with BMP-2.

AIM: To test the hypothesis that a synthetic hydroxyapatite/ß-tricalcium phosphate (HA/TCP) construct combined with polyethylene glycol (PEG) hydrogel including recombinant human bone morphogenetic proteins-2 (rhBMP-2) enhances new bone formation compared with bone morphogenetic proteins-2 (BMP-2) delivered using the HA/TCP construct alone. MATERIAL AND METHODS: Bilateral mandibular partial thickness 20 × 8 × 8 mm (L × W × H) alveolar defects were surgically created in the edentulated posterior mandible in 18 female minipigs. Randomized into two groups of nine animals each, the alveolar defects either received HA/TCP or HA/TCP/PEG with or without BMP-2 (105 µg/defect) in contra-lateral sites using a split-mouth design. Primary outcome, bone density (%) within four regions of interest, was evaluated following a 4-week healing interval when the animals were killed for histometric analysis. RESULTS: Bone morphogenetic proteins-2 loaded onto HA/TCP constructs significantly enhanced new bone formation compared with HA/TCP controls. Adding PEG apparently obstructed BMP-2 induced bone formation. CONCLUSION: Polyethylene glycol compromises the osteogenic effect of BMP-2.

Successful Nd:YAG laser therapy for hair removal in the oral cavity after plastic reconstruction using hairy donor sites.

BACKGROUND: Concepts of reconstruction of intraoral structures may often include the transfer of flaps composed of external skin with hairs. Given that intraoral hair growth following myocutaneous flaps can cause discomfort, there is a need for effective treatments to relieve cancer patients of these symptoms. OBJECTIVE: To describe the successful epilation of hairy intraoral flaps using Nd:YAG laser emitting a wavelength of 1,064 nm. METHODS: We performed an interdisciplinary prospective clinical study with 9 patients suffering from intraoral hair growth after plastic reconstruction of a hairy donor site due to cancer. Eight male and one female patients were treated with 1-4 sessions of Nd:YAG laser at 5-15-week intervals. RESULTS: Laser treatment resulted in effective hair reduction in 8/9 patients regardless of flap type. In 5/9 patients a hair clearance of >90% could be achieved, whereas laser treatment was ineffective in one male with white hair. Patients were very satisfied with the outcome and no side effects could be observed. CONCLUSION: Nd:YAG laser therapy appears to be a successful therapeutic option for patients suffering from growth of dark hair in the oral cavity after plastic reconstruction using a hairy donor site.

Altered monocyte activation markers in Tourette's syndrome: a case-control study.

BACKGROUND: Infections and immunological processes are likely to be involved in the pathogenesis of Tourette's syndrome (TS). To determine possible common underlying immunological mechanisms, we focused on innate immunity and studied markers of inflammation, monocytes, and monocyte-derived cytokines. METHODS: In a cross-sectional study, we used current methods to determine the number of monocytes and levels of C-reactive protein (CRP) in 46 children, adolescents, and adult patients suffering from TS and in 43 healthy controls matched for age and sex. Tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6), soluble CD14 (sCD14), IL1-receptor antagonist (IL1-ra), and serum neopterin were detected by immunoassays. RESULTS: We found that CRP and neopterin levels and the number of monocytes were significantly higher in TS patients than in healthy controls. Serum concentrations of TNF-alpha, sIL1-ra, and sCD14 were significantly lower in TS patients. All measured values were within normal ranges and often close to detection limits. CONCLUSIONS: The present results point to a monocyte dysregulation in TS. This possible dysbalance in innate immunity could predispose to infections or autoimmune reactions.

Monoclonal antibodies for the immunotherapy of solid tumours.

More than 80% of all cancers are caused by solid malignancies. More than 90% of these tumours are of epithelial origin. The main principles in tumour treatment are surgery, radiotherapy, chemotherapy or combinations of these. Complete surgical removal of the tumour is the most effective therapy for solid malignancies. Recent advances in early cancer detection led to a higher rate of resectable primary tumours and therefore prognosis will be determined especially by metastasis. Even in early stages some tumour cells may disseminate for example into the bone marrow. If these occult metastasis get evident they are mostly incurable by surgery and often highly resistant to chemotherapy. Developing new therapeutic agents to destroy these resting cells is a major challenge for the improvement of cancer therapy in the future. Advances to reach these goals were made in immunological therapies with monoclonal antibodies (MABs). These MABs are for example directed against tumor-associated antigens (TAAs). By binding selectively on tumor cells they can activate immunological effector mechanisms, e.g. antibody dependent cell cytotoxicity (ADCC) or complement mediated lysis. Other mechanisms are the blocking of inhibiting molecules to re-activate anergic tumor infiltrating lymphocytes (TILs). Furthermore growing tumours depend on oxygen supply, i.e. on effective neovascularisation. Antibodies against VEGF are able to inhibit neovascularisation and are therefore used successfully in tumour therapy.

Accuracy of fit of 3-to-3 retainers after adhesive fixation using a neodymium-iron-boron magnet chain.

BACKGROUND AND OBJECTIVE: Six-base retainers are often used to prevent tertiary crowding. To minimize shear stress on the retainer, these should be fitted as precisely as possible. The aim of this study was to compare the accuracy of fit of 6-base retainers after adhesive fixation using a neodymium-iron-boron (NdFeB) magnet chain or a resin positioning aid. MATERIALS AND METHODS: A 6-base retainer was prepared for 40 pseudo-anonymous lower jaw models (2 × 20). Temporary fixation was performed with a resin positioning aid or a NdFeB magnet chain. The adhesive fixation of the retainers was randomized. The area and vertical distance between wire and teeth were determined by histomorphometry. The results were evaluated by ANOVA. The probability of error was specified as 5%. RESULTS: With the magnetic chain method, the area was significantly smaller than with the positioning aid (p = 0.0125). The difference between the two methods was ca. 1.3 mm². Moreover, the measured distance was affected significantly by the method used (p < 0.0002). With the magnet chain, the distance between the tooth and retainer was reduced by a mean of 0.05 mm. Conclusion. The fit of the retainer wire improves somewhat when the NdFeB magnet chain is used even though the wire is not as precisely positioned as it would be with a custom-made positioning aid.

Quality of life in school-age children with orofacial clefts and their families.

Nonsyndromic orofacial clefts might affect family functioning and probably reduce the quality of life in school-age children and their parents. One hundred seventy consecutive children with orofacial clefts between 8 and 12 years and their families were asked to answer the Impact on Family Scale and KINDL. The results were compared with the quality of life in an age- and sex-matched group of unaffected schoolchildren. One hundred thirty-two families participated in this study. Family functioning was found superior in families with children with cleft lip than in families with children with cleft palate only or cleft lip and palate. Sex had no significant influence on family functioning. The quality of life in schoolchildren with orofacial clefts was found superior to the control group. Reductions were observed in children with cleft lip and palate in the dimensions "family" and "friends," indicating problems in the social field. Boys with orofacial clefts experienced a lower quality of life than girls. Children with cleft lip and palate and cleft palate only experienced a lower quality of life than children with cleft lip. Even years after successful cleft reconstruction, coping and mastering the diagnosis of orofacial cleft is a relevant concern for affected families. Several limitations of the quality of life in schoolchildren were identified, mostly affecting their social role.

Imaging of the midpalatal suture in a porcine model: flat-panel volume computed tomography compared with multislice computed tomography.

OBJECTIVE: An investigation was conducted to compare the image quality of prototype flat-panel volume computed tomography (fpVCT) and multislice computed tomography (MSCT) of suture structures. STUDY DESIGN: Bone samples were taken from the midpalatal suture of 5 young (16 weeks) and 5 old (200 weeks) Sus scrofa domestica and fixed in formalin solution. An fpVCT prototype and an MSCT were used to obtain images of the specimens. The facial reformations were assessed by 4 observers using a 1 (excellent) to 5 (poor) rating scale for the weighted criteria visualization of the suture structure. A linear mixed model was used for statistical analysis. Results with P < .05 were considered to be statistically significant. RESULTS: The visualization of the suture of young specimens was significantly better than that of older animals (P < .001). The visualization of the suture with fpVCT was significantly better than that with MSCT (P < .001). CONCLUSION: Compared with MSCT, fpVCT produces superior results in the visualization of the midpalatal suture in a Sus scrofa domestica model.

Detection and size differentiation of simulated tooth root defects using flat-panel volume computerized tomography (fpVCT).

OBJECTIVE: Our aim was to evaluate the capacity of flat-panel volume computerized tomography (fpVCT) to enable the observer to detect and differentiate 3 different sizes of simulated tooth root defects in radiographs. STUDY DESIGN: Three simulated tooth root defects of different sizes and a defect-free area (160 sites in total) were randomly distributed on the buccal and lingual surface of 20 mandibular premolar roots of Sus scrofa domestica with round burs. For the imaging of the specimens, an fpVCT prototype was used. Findings were evaluated by 3 observers. RESULTS: Cavity 0 (no lesion) was correctly identified in 53%, cavity 1 in 69%, cavity 2 in 96%, and cavity 3 in 89%. Altogether, the simulated cavities were classified in a correct manner in 77%. The values were compared using receiver operating characteristic curves. The area under the curve (AUC) for cavity 0 versus the pooled results for cavities 1-3 was found to be 0.72. The AUC for the pooled results for 0-2 (no pathologic impact) versus cavity 3 (potential pathologic impact) was 0.94. There was no significant dependence of the results on the observer (P = .37). Results with P < .05 were considered to be significant. CONCLUSIONS: Flat-panel volume computerized tomography, which is currently used only as a research tool, has a high potential in detection and differentiation at an early stage of external root resorption cavities with pathologic relevance..

The use of a neodymium-iron-boron magnet device for positioning a multi-stranded wire retainer in lingual retention--a pilot study in humans.

The aim of this study was to evaluate the time requirement of a newly developed device made of neodymium-iron-boron (NdFeB) magnets for positioning a multi-stranded, canine-to-canine retainer during bonding compared with dental floss and a transfer tray. Forty-five patients aged between 12 and 33 years (26 male, 19 female) previously treated with fixed appliances were enrolled in the study. The patients were randomly allocated to three groups (15 per group). For each group a mandibular canine-to-canine retainer of 0.018 inch Dentaflex multi-stranded wire (Dentaurum) was prefabricated for each patient on a cast. The bonding procedure was identical, except for the method of positioning the wire during adhesive fixation: group A dental floss, group B a small prefabricated transfer tray of dental resin and group C the NdFeB magnet device. For each group, the time required for the complete bonding process was measured. Kruskal-Wallis and Wilcoxon-Mann-Whitney tests were used for group and pairwise comparisons, respectively. The three methods required statistically significant different times (P < 0.001). The Wilcoxon-Mann-Whitney test revealed that wire positioning with the magnet device was significantly faster [4.98 minutes; standard deviation (SD) 0.68 minutes] than with dental floss (7.65 minutes, SD 1.14 minutes; P = 0.0001) or with transfer tray (5.75 minutes, SD 0.57 minutes; P = 0.001). The NdFeB magnet device is a timesaving appliance for positioning a multi-stranded, canine-to-canine retainer during bonding when compared with dental floss and an individually prefabricated transfer tray.

Quality of life and family functioning in children with nonsyndromic orofacial clefts at preschool ages.

Children with orofacial clefts (OFC) at preschool ages may have to tolerate psychosocial disadvantages due to their altered speech and facial appearance probably affecting their quality of life (QoL) and family functioning. In 147 children with OFC aged between 5 and 6 years and their families, the QoL and family functioning were analyzed using the KINDL questionnaire for measuring health-related QoL in children and impact on family scale. The KINDL scores were lowest in the dimension self-esteem. In all dimensions, the KINDL scores of children were higher than those of the parents suggesting a superior QoL than the caregivers estimated (P<0.001). In affected families, the impact on family scale dimensions personal impact and impact on coping strategies were found highest. Families having children with isolated cleft lip or cleft lip and palate had higher impacts on coping strategies when compared with children having isolated cleft palate (P<0.041). The impact for siblings (P<0.02) was found highest in patients with cleft lip and palate. In all examined dimensions, children with OFC perceived a higher QoL than their caregivers expected. However, self-esteem seems to be problematic in all types of OFC and in both genders. Knowledge of potential impacts related to the type of cleft and the gender of the patient will probably facilitate health care professionals to identify children and families at high risk to experience a reduced QoL and may help to offer specific support and treatment strategies.

CPA6, FMO2, LGI1, SIAT1 and TNC are differentially expressed in early- and late-stage oral squamous cell carcinoma--a pilot study.

To identify novel genes that could be involved in oncogenesis of oral squamous cell carcinoma a microarray-based gene-expression analysis was performed using tumour samples from patients with low-stage (n=4) and high-stage (n=4) disease in a pilot study. Genes (601) were found to be significantly regulated in cancer tissue compared to adjacent intraindividual mucosa controls. Genes (25) were identified with differences in their regulation comparing samples from early-stage cancer with those from advanced disease. The gene expression pattern of 5 of 7 genes examined by real-time-PCR verified the results received from the microarray-experiment. Among these, FMO2, CPA6, TNC and SIAT1 were significantly upregulated in early disease stages. LGI1 gene expression was significantly enhanced in normal adjacent mucosa of patients with early-stage disease without showing a differential expression in carcinoma biopsies. With this pilot study several novel genes were identified, which could be related to early and late stage disease. Hypotheses from these findings are discussed and have to be confirmed in a larger study sample.